DNA Segments Sensitive to Single-Strand-Specific Nucleases Are Present in Chromatin of Mitotic Cells

It was observed before that DNAin situin chromatin of mitotic cells is more sensitive to denaturation than DNA in chromatin of interphase cells. DNA sensitivity to denaturation, in these studies, was analyzed by exposing cells to heat or acid and using acridine orange (AO), the metachromatic fluorochrome which can differentially stain double-stranded (ds) vs single-stranded (ss) nucleic acids, as a marker of the degree of DNA denaturation. However, without prior cell treatment with heat or acid no presence of single-stranded DNA in either mitotic or interphase cells was detected by this assay. In the present experiments we demonstrate that DNAin situin mitotic cells, without any prior treatment that can induce DNA denaturation, is sensitive to ss-specific S1 and mung bean nucleases. Incubation of permeabilized human T cell leukemic MOLT-4, promyelocytic HL-60, histiomonocytic lymphoma U937 cells, or normal PHA-stimulated lymphocytes with S1 or mung bean nucleases generated extensive DNA breakage in mitotic cells. DNA strand breaks were detected using fluorochrome-labeled triphosphonucleotides in the reaction catalyzed by exogenous terminal deoxynucleotidyl transferase. Under identical conditions of the cells’ exposure to ss-specific nucleases, DNA breakage in interphase cells was of an order of magnitude less extensive compared to mitotic cells. The data indicate that segments of DNA in mitotic chromosomes, in contrast to interphase cells, may be in a conformation which is sensitive to ss nucleases. This may be a reflection of the differences in the torsional stress of DNA loops between interphase and mitotic chromatin. Namely, greater stress in mitotic loops may lead to formation of the hairpin-loop structures by inverted repeats; such structures are sensitive to ss nucleases. The present method of detection of such segments appears to be more sensitive than the use of AO. The identification of mitotic cells based on sensitivity of their DNA to ss nucleases provides an additional method for their quantification by flow cytometry.     

丝裂素活化蛋白激酶参与内皮素刺激的兔主动脉平滑肌细胞增生

内皮素（ｅｎｄｏｔｈｅｌｉｎ,ＥＴ）是已知的体内活性最强的缩血管物质,其缩血管作用由Ｇ蛋白偶联受体所介导。但ＥＴ强大的促血管平滑肌细胞（ＶＳＭＣ）增生效应的机理尚未完全阐明。本研究选用培养的兔胸主动脉ＶＳＭＣ,探讨丝裂素活化蛋白激酶（ＭＡＰＫ）在ＥＴ促细胞增生中的作用。结果表明：ＥＴ－１呈时间和浓度依赖性地促进细胞摄取￣３Ｈ－ＴｄＲ和激活ＭＡＰＫ,此作用可被蛋白激酶Ｃ（ｐｒｏｔｅｉｎｋｉｎａｓｅＣ,ＰＫＣ）抑制剂Ｓｔａｕｒｏｓｐｏｒｉｎｅ（ＳＴＰ）,Ｈ－７和ＥＴ＿Ａ受体拮抗剂ＢＱ１２３所抑制,但不被酪氨酸激酶抑制剂ＨｅｒｂｉｍｙｃｉｎＡ（Ｈｅｒｂ）所抑制,用ＰＫＣ激动剂ＰＭＡ（Ｐｈｏｒｂｏｌｍｙｒｉｓｔａｔｅａｃｅｔａｔｅ）预处理ＶＳＭＣ,使其ＰＫＣ活性下调,可显著减弱ＥＴ－１对ＭＡＰＫ的激活能力。本结果提示：（１）ＭＡＰＫ参与ＥＴ－１所致的ＶＳＭＣ增生;（２）ＥＴ－１促细胞增生与激活ＭＡＰＫ的作用是由ＥＴ＿Ａ受体和ＰＫＣ介导的。     

Phloem Mobility of Boron is Species Dependent: Evidence for Phloem Mobility in Sorbitol-rich Species

Boron is generally considered to be phloem immobile or to haveonly limited phloem mobility in higher plants. Evidence suggests,however, that B may be mobile in some species within thePyrus,Malus andPrunusgenera. These genera utilize sorbitol as a primarytranslocated photosynthate and it has been clearly demonstratedthat B forms stable complexes with sorbitolin vitro.In the researchpresented here we demonstrate, further, that B is freely phloemmobile inPyrus, MalusandPrunusspecies and suggest that thisis mediated by the formation and transport of B-sorbitol complexes. The pattern of B distribution within shoot organs and the translocationof foliar-applied, isotopically-enriched¹⁰B was studied in sixtree species. Results demonstrate that in species in which sorbitolis a major sugar (sorbitol-rich), B is freely mobile while inspecies that produce little or no sorbitol (sorbitol-poor) Bis largely immobile. The sorbitol-rich species used here werealmond [Prunus amygdalusB. syn.P. dulcis(Mill.)], apple (MalusdomesticaB.) and nectarine (Prunus persicaL. B. var.nectarinaM.),sorbitol-poor species included fig (Ficus caricaL.), pistachio(Pistacia veraL.) and walnut (Juglans regiaL.). In sorbitol-richspecies foliar applied¹⁰B was transported from the treated leavesto adjacent fruit and specifically to the fruit tissues (hull,shell or kernel) that developed during the experimental period.Whereas, foliar-applied¹⁰B was rapidly translocated out of leaves,only a small percentage of the¹¹B present in the leaf at thetime of foliar application was retranslocated. In sorbitol-richspecies, B concentrations differed only slightly between oldand young leaves while fruit tissue had significantly greaterB concentrations than leaves. In contrast, sorbitol-poor specieshad significantly higher B concentrations in older leaves thanyoung leaves while fruit tissue had the lowest B concentration.This occurred irrespective the source of plant B (soil, solutionor foliar-applied). In a subsequent experiment the growth ofapple trees in solutions free of applied B was maintained solelyby foliar applications of B to mature leaves. These resultsindicate that B is mobile in species that produce significantamounts of sorbitol. We propose that the mobility of B in thesespecies is mediated by the formation of B-sorbitol complexes. Almond; Prunus amygdalus ; apple; Malus domestica; nectarine; Prunus persica; fig; Ficus carica; pistachio; Pistacia vera; walnut; Juglans regia; boron; phloem mobility; deficiency; toxicity; inductively coupled plasma-mass; spectrometer     

以异源四倍体体细胞杂种为父本杂交培育三倍体柑桔植株的研究

以 3个柑桔原生质体融合而来的四倍体体细胞杂种为父本 ,与二倍体单胚性种柚子 (Citrusgrandis)以及单多胚混合型品种“华农本地早”桔 (C.reticulata)有性杂交 ,授粉后 90 d,发现种子干瘪 ,大部分种子的胚败育。将干瘪种子在 MT附加 1mg/L GA3 或 50 0 mg/L麦芽浸出物的培养基中 ,经培养抢救 ,有 2 5.6%的种子萌发成苗或继续进行胚的生长 ,后者进一步诱导能形成丛芽 ,经试管嫁接或诱导生根形成完整植株。共获得 6个组合 73棵完整植株 ,染色体数检查表明 ,2 0株为三倍体 (2 n=3x=2 7) ,32株为二倍体 (2 n=2 x=18) ,8株为非整倍体 ,其它 13株还有待于进一步检查。     

小檗科鬼臼亚科植物的核型研究

本文首次报道了中华山荷叶与川八角莲的核型,分别为Ｋ（２ｎ）＝１２＝８ｍ（４ＳＡＴ）＋２ｓｔ＋２ｔ及Ｋ（２ｎ）＝１２＝４ｍ（２ＳＡＴ）十４ｓｍ＋２ｓｔ（２ＳＡＴ）＋２ｔ,核型类型均为ＺＡ型。本文报道的桃儿七及八角莲的核型与前人的结果有一定差异,前者为：Ｋ（２ｎ）＝１２＝６ｍ（４ＳＡＴ）＋２ｓｍ＋２ｓｔ＋２ｔ,２Ｂ型,后者为Ｋ（２ｎ）＝１２＝８ｍ（２ＳＡＴ）＋２ｓｔ（２ＳＡＴ）＋２ｔ,为２Ａ型。本文分析了小檗科鬼臼亚科４个属共７种植物的核型,结果是该类植物的核型极为相似,染色体数目均为２ｎ＝１２,由８条ｍ或ｓｍ,２条ｓｔ以及２条ｔ染色体组成。核型的相似性反映了这类植物的亲缘关系,这４个属的植物是一个自然类群。但随着系统发育,核型的不对称性有所增加,其中以山荷叶属最为对称,八角莲属居中,桃儿七属与足叶草属最不对称。笔者认为,核型上的高度相似是该类植物在系统发育上不发达,属内种类稀少,通常为寡种属或单种属的重要原因。     

懒猴属的核糖体DNA变异及其种间分化关系

用１５种限制性内切酶和人２８Ｓ、１８ＳｒＤＮＡ探针构建了懒猴属各物种核糖体ＤＮＡ重复单位的限制性内切酶图谱。在进化速率较高的非转录间隔区,在大、中、小懒猴中分别定位了２３、２４、２４个酶切位点。大懒猴与中懒猴有１２个位点不同,与小懒猴有１４个位点不同,而中、小懒猴间则只有一个位点的差异。经过计算,大懒猴与中懒猴的遗传距离值为１２．６５％,与小懒猴的差异为１４．２４％,中、小懒猴间的差异则仅为０．７     

云南姬鼠的蛋白多态性及其遗传分化关系

本文采用蛋白电泳技术对来源于云南省若干地区的姬鼠属（Ａｐｏｄｅｍｕｓ）的３种姬鼠──高山姬鼠（Ａ．ｃｈｅｖｒｉｅｒｉ）８只,中华姬鼠（Ａ．ｄｒａｃｏ）３只和大耳姬鼠（Ａ．ｌａｔｒｏｎｕｍ）１只,以及作为外群的同科的绒鼠属的大绒鼠（Ｈａｐａｌｏｍｙｓｄｅｌａｌｏｒｉ）３只进行了分析。共检测遗传座位２７个,发现２１个座位存在多态性。根据蛋白多态的数据对研究对象进行遗传分化关系的探讨,用系统分析软件ＰＨＹＬＩＰ计算它们之间的分化关系,得到了一棵无根系统树。结果表明,作为外群的大绒鼠明显不同于其它３种姬鼠而聚在最外面。８只高山姬鼠个体汇聚成独立的一支,中华姬鼠的３个个体也聚成一支,但大耳姬鼠却聚在中华姬鼠一支中,因此我们认为大耳姬鼠同中华姬鼠的分化时间可能比较晚近。     

High-performance liquid chromatographic method for the determination of a novel thymidylate synthase inhibitor, AG 331, in human serum

AG 331 is a novel thymidylate synthase inhibitor currently in Phase I clinical trial. To determine the pharmacokinetic parameters of AG 331 in human subjects, a suitable analytical method was developed using high-performance liquid chromatography. Serum and urine samples were prepared using both solid-phase extraction and solvent extraction. Either 4,4′-diaminodiphenyl sulfone or benz[cd]indole-2(1H)-one were used as internal standards for the method. A reversed-phase C₁₈ analytical column completely resolved the drug and internal standard peaks from non-specific substances present in biological matrix. The method was validated for precision, accuracy, and reproducibility in serum and was linear over a concentration range of 50–2000 ng/ml, with a limit of detection of 20.0 ng/ml and a quantifiable limit of 50 ng/ml.     

地膜条件下土壤生态因子与玉米产量的关系

研究干旱地区地膜条件下,土壤生态因子对玉米产量的影响．结果表明,最佳种植模式为作物距地膜中心线２１．３—３３．２Ｃｍ,施纯Ｎ＋Ｐ为５９８—７４３ｋｇ·ｈａ－１,玉米产量为１４４９２≤Ｙ佳≤１６５９ｋｇ·ｈ－１．该种植模式有利于提高作物产量及残膜回收．     

大兴安岭林区针叶林的生长方程及火灾林木死亡率

本文用随机模拟的方法建立了北方针叶林的生长方程及其受到火灾侵害后的林木死亡率,定量地研究了火灾对这种林分变动的影响．